Dynamic Flux Research Articles

Dynamic flux balance analysis of high cell density fed-batch culture of Escherichia coli BL21 (DE3) with mass spectrometry-based spent media analysis.

Dynamic flux balance analysis (FBA) allows estimation of intracellular reaction rates using organism-specific genome-scale metabolic models (GSMM) and by assuming instantaneous pseudo-steady states for processes that are inherently dynamic. This technique is well-suited for industrial bioprocesses employing complex media characterized by a hierarchy of substrate uptake and product secretion. However, knowledge of exchange rates of many components of the media would be required to obtain meaningful results. Here, we performed spent media analysis using mass spectrometry coupled with liquid and gas chromatography for a fed-batch, high-cell density cultivation of Escherichia coli BL21(DE3) expressing a recombinant protein. Time course measurements thus obtained for 246 metabolites were converted to instantaneous exchange rates. These were then used as constraints for dynamic FBA using a previously reported GSMM, thus providing insights into how the flux map evolves through the process. Changes in tri-carboxylic acid cycle fluxes correlated with the increased demand for energy during recombinant protein production. The results show how amino acids act as hubs for the synthesis of other cellular metabolites. Our results provide a deeper understanding of an industrial bioprocess and will have implications in further optimizing the process.

Stratification of dissolved organic matter in the upper 5000 m water column in the western Pacific Ocean

Marine dissolved organic matter (DOM) is a principal reservoir involved in biogeochemical cycles and exerts a pivotal influence on global carbon flux dynamics. In this study, excitation-emission matrix fluorescence spectroscopy combined with parallel factor analysis (EEM-PARAFAC) was conducted on 230 DOM samples collected from 21 sites between February and April 2022 in the Western Pacific Ocean (WPO). We identified five distinct fluorescence peaks (peaks B, T, A, C, and M), predominantly protein-like and humic-like components. These findings, marked by significant differences (p < 0.01) in fluorescence intensities and spectral indices, characterized the transformation of DOM with ocean depth, illustrating a transition from active to recalcitrant forms. Additionally, random forest analysis (RFA) identified depth as a key factor influencing marine dissolved organic carbon (DOC), with a 32.59% importance value. Correlations between hydrological and fluorescent parameters underscored the complexity of DOM sources and influencing processes. Overall, this work broadens our understanding of DOM variability in the upper 5000 m of the WPO, enhancing our knowledge of the marine environment's role in the global carbon cycle.

Greenhouse gas emission flux (CO2, CH4, N2O) from marine shrimp (Litopenaeus vannamei) monoculture cultures in the Brazilian semi-arid region

Greenhouse gases (GHG) and their rising atmospheric concentrations have been one of the most important international subjects due to their huge impact on climate change (IPCC, 2022). Aquaculture is of crucial importance to the human food supply and nutrition since it is one of the major sectors providing animal protein, besides reducing the pressure on the fishery stocks from natural resources worldwide (Cai, Zhou, 2019; FAO, 2022). On the other hand, it represents a threat to the environment by generating waste. Thus, this research aims to evaluate the GHG flux dynamics in marine shrimp (Litopenaeus vannamei) monoculture cultures. The study was carried out in two aquaculture farms, both located in the Brazilian semi-arid region. In order to measure the production of greenhouse gases, three cultures of marine shrimp were monitored. The dynamics of the diffusive and ebullitive emission of CO2, CH4, and N2O were monitored in the three units during the daytime (6:00 AM) and nighttime (6:00 PM). The results showed the stocking density as the main regulator for ebullitive CO2 emissions. A decreasing trend in diffusive CO2 emissions was observed. Aeration played a crucial role in controlling the diffusive and ebullitive emissions of CH4 and N2O, effectively reducing their release. The diffusive CH4 emissions exceeded ebullitive emissions and the oxygen content suppressed the anoxic pathway for N2O, and the ponds acted as a sink for this gas. This research aims to evaluate the dynamics of greenhouse gas flow in marine shrimp (Litopenaeus vannamei) monoculture cultures in the Brazilian semi-arid region and to verify whether lower stocking densities emit less greenhouse gases when compared to higher densities.

Optimizing Fed-Batch Processes with Dynamic Control Flux Balance Analysis

Fed-batch processes are prevalent in biotechnological industries, but design of experiments often results in sub-optimal conditions due to incomplete solution space characterization. We employ a single-level dynamic control (DC) algorithm for dynamic flux balance analysis (dFBA), enhancing efficiency by reducing Karush-Kuhn-Tucker (KKT) condition constraints and adapting the algorithm for predicting optimal process length. In a growth-decoupled plasmid DNA production case study, we predict the optimal feeding profile and switching time between growth and production phase. Comparing our algorithm to its predecessor shows a speed-up of at least a factor of four. When the process length is part of the objective function the speed-up becomes considerably larger.

Characterization and drivers of water and carbon fluxes dynamics in dune ecosystems of the Horqin Sandy Land

Sandy regions constitute pivotal components of terrestrial ecosystems, exerting significant influences on global ecological equilibrium and security. This study meticulously explored water and carbon fluxes dynamics within a dune ecosystem in the Horqin Sandy Land throughout the growing seasons from 2013 to 2022 by employing an advanced eddy covariance system. The dynamic characteristics of these fluxes and their underlying driving forces were extensively analyzed, with a particular focus on the impact of precipitation. The main results are as follows: (1) During the growing seasons of 2015 and 2016, the dune ecosystem acted as a modest carbon source, while in 2013, 2014, and 2017– 2022, it transformed into a net carbon sink. Notably, the annual mean values of water use efficiency (WUE) and evapotranspiration (ET) were 5.16 gC·kg−1H2O and 255.4 mm, respectively. (2) The intensity, frequency, and temporal distribution of precipitation were found to significantly influence the carbon and water fluxes dynamics. Isolated minor precipitation events did not trigger substantial fluctuations, but substantial and prolonged precipitation events spanning multiple days or consecutive minor precipitation events resulted in notable assimilation delays. (3) Air temperature, soil temperature, and fractional vegetation cover (FVC) were found to be key factors influencing the carbon and water fluxes. Specifically, FVC exhibited a negative logarithmic correlation with net ecosystem CO2 exchange (NEE) and a power function relationship with WUE. (4) The interaction between carbon and water fluxes is exhibited by exponential increases in ecosystem respiration (Reco) and gross primary productivity (GPP) with WUE, while NEE displayed an exponential decrease in relation to WUE. These findings are of high significance in predicting the potential ramifications of climate change on the intricate carbon and water cycles, and enhance our understanding of ecosystem dynamics in sandy environments.

Mineralization and temperature sensitivity of soil organic matter pools of contrasting lability

AbstractUnderstanding the temperature sensitivity (Q10) of soil organic matter (SOM) decomposition is crucial to predict CO2 emissions and carbon (C) stocks under global warming. This study describes the decomposition and Q10 of four soil C pools: (1) very labile (glucose addition (GLU), representing root exudates), (2) labile (microbial turnover, MT), (3) potentially labile (primed C pool, PE), and (4) resistant (inherent soil C, RES). The soil (loamy Luvisol) was incubated for 4 and 144 days at five temperatures (0, 10, 20, 30, and 40°C) with or without 14C‐labeled glucose. The dynamics of CO2 fluxes were measured during short (0–4 days) and long (5–144 days) term incubations. Glucose was mineralized following a two‐pool exponential function. The half‐life of the decomposition of the GLU pool decreased by 3 times as temperature increased from 10 to 40 °C. The flux of unlabeled CO2 over 4 days reflects a strong contribution to the apparent priming especially at high temperature, which was due to the accelerated microbial biomass turnover. Accordingly, the CO2 flux increased during short‐term incubation and was dominated by the decomposition of labile SOM and microbial biomass turnover, whereas during the long‐term incubation, the CO2 was mainly released from the temperature‐stimulated decomposition of RES pool. The short‐term Q10 of the soil C pools decreased in the order: GLU (2.1) &gt; MT (1.8) &gt; PE (1.3) ≈ RES (1.6) over a few days (0–4 days), but the Q10 measured over the long‐term period (144 days) was in the range of 1.2 (PE) to 1.8 (RES) and decreased in the order RES &gt; MT &gt; PE &gt; GLU. In conclusion, CO2 emissions linearly increased with temperature in all pools over short‐ and long‐term incubation, except for the GLU pool during long‐term incubation. The Q10 strongly depends on the availability of C pools for microorganisms and decreases over time with the exhaustion of available substances in soil. This needs to be considered when estimating temperature effects on CO2 emissions and C turnover in soil.

MAGNETOTHERMAL PROPERTIES OF COPPER FERRITE

In this work, we carried out the directed synthesis of copper ferrite (CuFe2O4) by co-precipitation from solutions with an excess of alkali at a temperature of 373 К. The methods of scanning electron microscopy and X-ray phase analysis were used to study the crystal structure and surface morphology of copper ferrite. The ratio of the characteristic peaks of the diffractogram was carried out in accordance with the JCPDS database. It is shown that copper (II) ferrite crystallizes in the structure of the cubic spinel. Microphotographs of the synthesized copper ferrite samples at different magnifications are given. It is established that ferrite particles consist of conglomerates of crystals with different sizes of individual grains. Their small scatter in size is observed. The average particle size is in the range of 100-200 nm. Using a dynamic heat flux differential scanning calorimeter, the temperature dependence of the heat capacity of copper ferrite samples was obtained in the temperature range of 273–373 K. Using an original microcalorimetric setup in the temperature range of 288–346 K and with a change in the magnetic field induction from 0 to 1.0 T, thermodynamic characteristics were obtained - the magnetocaloric effect (MCE) and the change in entropy (ΔS) during the magnetization of copper (II) ferrite. It was found that with an increase in the magnitude of the magnetic field, the magnitude of the MCE increases and decreases with increasing temperature. It was found that the temperature dependences of the magnetocaloric effect and specific heat have an extreme character. In the region of room temperatures (308–315 K), a maximum of the magnetocaloric effect and a minimum of the specific heat are observed. This previously unknown anomalous behavior of the magneto-thermal properties of copper II ferrite was discovered for the first time.

Hyperpolarized Carbon-13 Magnetic Resonance Imaging: Technical Considerations and Clinical Applications.

Hyperpolarized (HP) carbon-13 (13C) MRI represents an innovative approach for noninvasive, real-time assessment of dynamic metabolic flux, with potential integration into routine clinical MRI. The use of [1-13C]pyruvate as a probe and its conversion to [1-13C]lactate constitute an extensively explored metabolic pathway. This review comprehensively outlines the establishment of HP 13C-MRI, covering multidisciplinary team collaboration, hardware prerequisites, probe preparation, hyperpolarization techniques, imaging acquisition, and data analysis. This article discusses the clinical applications of HP 13C-MRI across various anatomical domains, including the brain, heart, skeletal muscle, breast, liver, kidney, pancreas, and prostate. Each section highlights the specific applications and findings pertinent to these regions, emphasizing the potential versatility of HP 13C-MRI in diverse clinical contexts. This review serves as a comprehensive update, bridging technical aspects with clinical applications and offering insights into the ongoing advancements in HP 13C-MRI.

Sensorless Control of Electrically Excited Synchronous Machines Considering Magnetic Saturation and Flux Linkage Dynamics for Automotive Applications

For electrically excited synchronous machines (EESMs) used in automotive applications, the flux linkages are non-linear and the magnetic saturation plays an important role. The coupling between d-axis and exciter axis in EESMs introduces a significant fluctuation of the flux linkages due to current changes. To cope with these problems, a novel observer incorporating flux linkage saturation and dynamics caused by the coupling is proposed. The flux is not regarded as linear, but the absolute and differential inductances are used in the observer. Moreover, the current dynamics induced flux variations are compensated for. As the system is nonlinear and time-varying, Lyapunov's method is used to prove the stability of the system and to derive a speed-adaptive algorithm. The experimental results on an EESM test bench confirm the effectiveness of the proposed method.

Probing interspecies metabolic interactions within a synthetic binary microbiome using genome-scale modeling.

Aim: Metabolic interactions within a microbial community play a key role in determining the structure, function, and composition of the community. However, due to the complexity and intractability of natural microbiomes, limited knowledge is available on interspecies interactions within a community. In this work, using a binary synthetic microbiome, a methanotroph-photoautotroph (M-P) coculture, as the model system, we examined different genome-scale metabolic modeling (GEM) approaches to gain a better understanding of the metabolic interactions within the coculture, how they contribute to the enhanced growth observed in the coculture, and how they evolve over time. Methods: Using batch growth data of the model M-P coculture, we compared three GEM approaches for microbial communities. Two of the methods are existing approaches: SteadyCom, a steady state GEM, and dynamic flux balance analysis (DFBA) Lab, a dynamic GEM. We also proposed an improved dynamic GEM approach, DynamiCom, for the M-P coculture. Results: SteadyCom can predict the metabolic interactions within the coculture but not their dynamic evolutions; DFBA Lab can predict the dynamics of the coculture but cannot identify interspecies interactions. DynamiCom was able to identify the cross-fed metabolite within the coculture, as well as predict the evolution of the interspecies interactions over time. Conclusion: A new dynamic GEM approach, DynamiCom, was developed for a model M-P coculture. Constrained by the predictions from a validated kinetic model, DynamiCom consistently predicted the top metabolites being exchanged in the M-P coculture, as well as the establishment of the mutualistic N-exchange between the methanotroph and cyanobacteria. The interspecies interactions and their dynamic evolution predicted by DynamiCom are supported by ample evidence in the literature on methanotroph, cyanobacteria, and other cyanobacteria-heterotroph cocultures.

Flux dynamics of C-5 amino acid precursors in fed-batch cultures of Streptomyces clavuligerus during clavulanic acid biosynthesis

Clavulanic acid (CA) is a well-known β-lactamase inhibitor that is mainly obtained from submerged cultures of Streptomyces clavuligerus. Dynamic genome-scale in silico studies were performed to gain insights into the intracellular metabolic fluxes of C-5 precursors during the cultivation of wild-type S. clavuligerus in batch and fed-batch operation. A preliminary literature screening was conducted for media selection and determination of culture conditions for further cultivation in a 5-L bioreactor. Simulations provided insights into the metabolism of C-5 precursors in CA biosynthesis. In addition, carbon utilization for CA biosynthesis was assessed in terms of the ratio of total carbon used in CA biosynthesis to the total carbon influx in batch and feed media. Based on simulation results, we proposed a modification of the glycerol-sucrose-proline-glutamate medium for fed-batch cultivation to improve the carbon utilization for CA biosynthesis. The proposed fed-batch scenario achieved higher specific CA concentration at lower biomass production, indicating better carbon utilization for its synthesis. The dynamic in silico fluxes suggests that metabolic fluxes in this scenario would be stable, favoring a longer stage of continued antibiotic secretion.

A Greenhouse Gas Budget for Mexico During 2000–2019

AbstractApplication of the best available science to improve quantification of greenhouse gas (GHG) emissions at regional and national scales is key to climate action. Here, we present a two‐decade (2000–2019) GHG (CO2, CH4, and N2O) budget for Mexico derived from multiple products. Data from the National GHG Inventory, global observations, and the scientific literature were compared to identify knowledge gaps on GHG flux dynamics and discrepancies among estimates. Total mean annual GHG emissions were estimated at 695–910 TgCO2‐eq year−1 over these two decades, with 70% of the emissions attributable to CO2, 23% to CH4, and 5% to N2O (2% to other gases). When divided by sectors, we found agreement across emission estimates from various sources for fossil fuels, cattle, agriculture, and waste for all GHGs. However, considerable discrepancies were identified in the fluxes from terrestrial ecosystems. The disagreement was particularly large for the land CO2 sink, where net biome production estimations from the national inventory were double those from any other observational product. Extensive knowledge gaps exist, mainly related to aquatic systems (e.g., outgassing in rivers) and the lateral fluxes (e.g., wood trade). In addition, limited information is available on CH4 emissions from wetlands and soil CH4 consumption. We expect these results to guide future research to reduce estimation uncertainties and fill the information gaps across Mexico.

The impact of fast-growing eucalypt plantations on C emissions in tropical soil: effect of belowground and aboveground C inputs.

Planted forest soils can have great potential for CO2-C sequestration, mainly due to belowground C inputs, which impact deep soil C (DSC) accumulation. However, there are still gaps in understanding the CO2 emission dynamics in eucalypt plantations. Therefore, we used isotopic techniques to investigate the dynamics of the soil surface CO2-C flux and CO2-C concentration with depth for a eucalypt plantation influenced by different C inputs (above- and belowground). The gas evaluations were carried in depth the root to valuation of root priming effect (RPE) was calculated. In addition, measurements of the plant (C-fine root and C-litterfall) and soil (total organic carbon - TOC, total nitrogen - TN, soil moisture - SM, and soil temperature - ST) were performed. After planting the eucalypt trees, there was an increase in the soil surface CO2-C flux with plant growth. Root growth contributed greatly to the soil surface CO2-C flux, promoting greater surface RPE over time. In comparison to the other factors, SM had a greater influence on litterfall decomposition and root respiration. It was not possible to detect losses in TOC and TN in the different soil layers for the 31-month-old eucalypt. However, the 40-month-old eucalypt showed a positive RPE with depth, indicating possible replacement of DSC ("old C") by rhizodeposition-C ("new C") in the soil. Thus, in eucalyptus plantations, aboveground plant growth influences CO2 emissions on the soil surface, while root growth and activity influence C in deeper soil layers. This information indicates the need for future changes in forest management, with a view to reducing CO2 emissions.

Multiphysics Simulation of a Novel Self-Adaptive Chip Cooling with a Temperature-Regulated Metal Pillar Array in Microfluidic Channels

Conventional liquid cooling techniques may provide effective chip cooling but at the expense of high pumping power consumption. Considering that there is dynamic heat load in practice, a self-adaptive cooling technique is desired to reduce operational costs while preserving inherent cooling effectiveness. In this work, a novel self-adaptive cooling strategy is presented to balance the thermal and flow efficiency in accordance with the dynamic thermal load, based on temperature-regulated movement of the metal pillar array in a microfluidic channel. With an illustrative device, the effectiveness of such a strategy is investigated using multiphysics modeling and simulation. As a case study, the device is considered to be initiated with a chip power of 5 W and an inlet coolant velocity of 0.3 m/s. It is shown that the temperature-regulated movement of the metal pillar heat sink will be activated rapidly and equilibrate within 30 s. Parts of the metal pillars immerse into the coolant flow, resulting in significantly improved heat transfer efficiency. The diminished thermal resistance leads to a reduction in chip temperature rise from 225 K (without structural adaptation) to 91.86 K (with structural adaption). Meanwhile, the immersion of metal pillars into the coolant also causes an increased flow resistance in the microfluidic channel (i.e., pressure drop increases from 859.27 Pa to 915.98 Pa). Nevertheless, the flow resistance decreases spontaneously when the working power of the chip decreases. Comprehensive simulation has demonstrated that the temperature-regulated structure works well under various conditions. Therefore, it is believed that the presented self-adaptive cooling strategy offers simple and cost-effective thermal management for modern electronics with dynamic heat fluxes.

The coupled effect of sediment resuspension and microbiota on phosphorus release and transformation in a simulated aquatic ecosystem

Plant microbial involvement in phosphorus (P) release and transformation during resuspension events in bottom aquatic systems remains poorly understood. Laboratory experiments were used in this study to explore the release and transformation of P from sediments and the microbiota involved. Dissolved and labile P were obtained by high-resolution dialysis and the diffusive gradients in thin films (DGT) technique, respectively. The bacterial community was investigated using a scanning electron microscope and 16S rRNA high throughput sequencing technique. Finally, the diffusion fluxes dynamics of phosphorus were calculated using the DGT-induced fluxes in sediments and soils (DIFS) model. These results showed that sustained disturbance can balance dissolved P concentrations in the resuspension layer (overlying water) and enhance the stability of P transformation and release processes. Notably, the bacterial community within the deeper sediment layer (Below 10 mm sediment depth) significantly influences P interception. Acidobacteria and Neobacteria were identified as the key bacterial genera responsible for phosphorus adsorption of plant roots at speeds of 150 rad/min (R150). In addition, as the sediment depths increased, a slower decrease in R curves fitted with the DIFS model, which indicated a continuous resupply process from solid phase to pore water, especially below R150. This study provides theoretical references for managing shallow lakes with frequent sediment resuspension.

Map to Compassion

This article introduces the Human Needs Map — a sense-making tool that helps orient our minds to human needs that drive and trigger us, disrupting relationships and creating conflict. Initially inspired from fieldwork of the Israeli-Palestinian peace process, the Human Needs Map has developed into a systems model that reveals the interior landscape of our Protective body/mind with needs and emotions in dynamic flux. The model provides a language to speak about the interconnectivity of needs through their synergies and tensions. Naming this emerging and coherent inter-dependency between needs and emotions offers a way to understand emotions as reasonable and provides more vectors to heal, dislodge or re-write narratives and beliefs, which perpetuate conflict. The implication lies not only at the individual level but also at the level of systems change to support the design of social and cultural structures more capable of taking needs and emotions into account. The present article traces the iterative process conceptualizing the model and discovering its internal patterns, followed by a discussion about the Needs system’s adaptive qualities and its role in creating and perpetuating Conflict, concluding with insights for Israeli-Palestinian peacebuilding.

Climate Factors Affect Above–Belowground Biomass Allocation in Broad-Leaved and Coniferous Forests by Regulating Soil Nutrients

The allocation of plant biomass above and below ground reflects their strategic resource utilization, crucial for understanding terrestrial carbon flux dynamics. In our comprehensive study, we analyzed biomass distribution patterns in 580 broadleaved and 345 coniferous forests across China from 2005 to 2020, aiming to discern spatial patterns and key drivers of belowground biomass proportion (BGBP) in these ecosystems. Our research revealed a consistent trend: BGBP decreases from northwest to southeast in both forest types. Importantly, coniferous forests exhibited significantly higher BGBP compared to broadleaved forests (p &lt; 0.001). While precipitation and soil nutrients primarily influenced biomass allocation in broadleaved forests, temperature and soil composition played a pivotal role in coniferous forests. Surprisingly, leaf traits had a negligible impact on BGBP (p &gt; 0.05). Climatic factors, such as temperature and rainfall, influenced biomass partitioning in both strata by altering soil nutrients, particularly soil pH. These findings provide valuable insights into understanding carbon sequestration dynamics in forest ecosystems and improving predictions of the future trajectory of this critical carbon cycle component.

Dynamic flux balance analysis of 1,3-propanediol production by clostridium butyricum fermentation.

To study the relationship between the yield of 1,3-propanediol (1,3-PDO) and the flux change of the Clostridium butyricum metabolic pathway, an optimized calculation method based on dynamic flux balance analysis was used by combining genome-scale flux balance analysis with a kinetic model. A more comprehensive and extensive metabolic pathway was obtained by optimization calculations. The primary extended branches include: the dihydroxyacetone node, which enters the pentose phosphate pathway; the α-oxoglutarate node, which has synthetic metabolic pathways for glutamic acid and amino acids; and the serine and homocysteine nodes, which produce cystathionine before homocysteine enters the methionine cycle pathway. According to the expanded metabolic network, the flux distribution of key nodes in the metabolic pathway and the relationship between the flux distribution ratio of nodes and the yield of 1,3-PDO were analyzed. At the dihydroxyacetone node, the flux of dihydroxyacetone converted to dihydroxyacetone phosphate was positively correlated with the yield of 1,3-PDO. As an important intermediate product, the flux change in the metabolic pathway of α-oxoglutarate reacting with amino acids to produce glutamic acid is positively correlated with the yield. When pyruvate was used as the central node to convert into lactic acid and α-oxoglutarate, the proportion of branch flux was negatively correlated with the yield of 1,3-PDO. These studies provide a theoretical basis for the optimization and further study of the metabolic pathway of C. butyricum.

Temporal Variability in Soil Greenhouse Gas Fluxes and Influencing Factors of a Primary Forest on the Eastern Qinghai-Tibetan Plateau

Soil greenhouse gas (GHG) fluxes relate to soil carbon and nitrogen budgets and have a significant impact on climate change. Nevertheless, the temporal variation and magnitude of the fluxes of all three major GHGs (CO2, CH4 and N2O) and their influencing factors have not been elucidated clearly in primary forests on the eastern Qinghai-Tibetan Plateau. Herein, field chamber GHG fluxes from May to November, soil microbial community and enzyme activity were analyzed in a fir-dominated (Abies fargesii var. faxoniana) primary forest. The emission rates of CO2 and N2O ranged between 64.69–243.22 mg CO2 m−2 h−1 and 1.69–5.46 ug N2O m−2 h−1, exhibiting a temporally unimodal pattern with a peak in July. The soil acted as a CH4 sink, and the uptake rate varied between 52.96 and 84.67 μg CH4 m−2 h−1 with the higher uptake rates in June and November. The temporal variation in the CO2 flux was significantly correlated with the geometric mean of enzyme activities, suggesting that the soil CO2 flux was determined by microbial activity rather than soil microbial biomass. The soil N2O flux was positively related to nitrate concentration with marginal significance, probably because N2O was a byproduct of nitrification and denitrification processes. The soil CH4 uptake was closely associated with methanotrophic biomass (18:1ω7c). The results highlight divergent temporal dynamics of GHG fluxes owing to different driving mechanisms and an important CH4 sink in the primary forest soil, helping to evaluate the carbon and nitrogen budgets of primary forests on the eastern Qinghai-Tibetan Plateau.

When do plant hydraulics matter in terrestrial biosphere modelling?

The ascent of water from the soil to the leaves of vascular plants, described by the study of plant hydraulics, regulates ecosystem responses to environmental forcing and recovery from stress periods. Several approaches to model plant hydraulics have been proposed. In this study, we introduce four different versions of plant hydraulics representations in the terrestrial biosphere model T&C to understand the significance of plant hydraulics to ecosystem functioning. We tested representations of plant hydraulics, investigating plant water capacitance, and long-term xylem damages following drought. The four models we tested were a combination of representations including or neglecting capacitance and including or neglecting xylem damage legacies. Using the models at six case studies spanning semiarid to tropical ecosystems, we quantify how plant xylem flow, plant water storage and long-term xylem damage can modulate overall water and carbon dynamics across multiple time scales. We show that as drought develops, models with plant hydraulics predict a slower onset of plant water stress, and a diurnal variability of water and carbon fluxes closer to observations. Plant water storage was found to be particularly important for the diurnal dynamics of water and carbon fluxes, with models that include plant water capacitance yielding better results. Models including permanent damage to conducting plant tissues show an additional significant drought legacy effect, limiting plant productivity during the recovery phase following major droughts. However, when considering ecosystem responses to the observed climate variability, plant hydraulic modules alone cannot significantly improve the overall model performance, even though they reproduce more realistic water and carbon dynamics. This opens new avenues for model development, explicitly linking plant hydraulics with additional ecosystem processes, such as plant phenology and improved carbon allocation algorithms.